Expression Of Eukaryon And Protokaryon Of H9N2Subtype Avian Influenza Virus HA Gene And Preparation Of Mono-specific Serum Against HA

The avian influenza epidemic and outbreak has caused great threat to the development ofpoultry industry and the public health security in recent years. Subtype H9N2, as thoughbelongs to lowly pathogenic awian influenza virus, has the capacity to spread widely and tocause human and animal diseases that may even give rise to pandemic which can’t be ignoredby us. Therefore, surveillance and control should be strengthened while numerous serotypesand high variability of influenza virus bring great challenge to the research.Avian influenza virus protein is consist of five peptides which are hemagglutinin,neuraminidase, Matrix Protein nucleoproteid and polymer. The hemagglutinin is aglycoprotein on the virus capsid and has the ability to agglutinate haemocytes of differentanimals. The virus absorb cells by the interactions between HA and cell receptors, which takeimportant roles in the membrane fusion prossess, and the highly varablity of HA contribute tothe antigenic variation. The HA protein has became worldwide research focus because it caninduce neutralizing antibodies to provide direct protection against AIV, which could play arole in the disease diagnosis and control.Firstly, the eukaryotic expressing plasmid of HA portein was successfully constructed,which provides a foundation for the researches. the HA gene of subtype H9N2AIV wascloned by RT-PCR into an expressing plasmid pcDNA3.1-A. Then the recombinant plasmidwas transfected into CEF cells with lipofectamine encapsuled to identify the expression of thetarget protein. At the same time, the prokaryotic expression vector was constructed, inducedin E. coli BL21by IPTG. Ternary form was affirmed by Comparing the quantity of the targetprotein encoded by different vectors, while no expression was detected when the HA proteinis integrated or divided by two parts. The white mouse were immuned by the HA protein andthe plasmids, the polyclonal antibodies were obtained, and the titer was determined by IFAassays. Comparing the character of those two kinds of antibodies, the result demonstrated that,both of those antibodies has high immunoreactivities and same titers. The research providedexperimental materials and datas to the further study of vaccine and diagnostic reagent ofAIV.